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An attempt to improve reproducibility of genotyping in RIKEN BRC: PCR-RFLP to Allele specific PCR

机译:尝试提高RIKEN BRC基因分型的可重复性:PCR-RFLP对等位基因特异性PCR

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摘要

RIKEN BRC has collected over 2,900 strains including various mutant mice. These include many mutant strains with point mutations, such as ENU mutant mice. We have used PCR- RFLP to genotype the mutant strain with point mutations so far. The reproducibility of PCR- RFLP method is not necessarily high enough, and is easily affected by reaction time for the enzyme digestion and the purity of DNA samples. With DNA polymerases of high tensibility and fidelity, we attempted to use allele-specific PCR (AS-PCR) to improve reproducibility of genotyping the mutant mice carrying point mutations.
机译:RIKEN BRC已收集了2900多种菌株,包括各种突变小鼠。这些包括许多具有点突变的突变株,例如ENU突变小鼠。到目前为止,我们已经使用PCR-RFLP对具有点突变的突变菌株进行基因分型。 PCR-RFLP方法的可重复性不一定足够高,并且容易受到酶消化反应时间和DNA样品纯度的影响。对于具有高张力和保真度的DNA聚合酶,我们尝试使用等位基因特异性PCR(AS-PCR)来提高携带点突变的突变小鼠的基因分型的可重复性。

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